title: Efficient electrophoretic method to remove neutral additives from protein solutions followed by mass spectrometry analysis.
abstract: A mass spectrometry (MS)-compatible, isoelectric point-based separation method for removal of neutral additives from protein solutions is described. The separation is performed by electrophoretic migration and trapping using a device referred to as membrane separated wells for isoelectric focusing and trapping (MSWIFT). Electrophoretic separation in the MSWIFT device is fast; the entire process can be carried out in a matter of minutes, and it does not require further sample cleanup prior to MS analysis. Proof-of-concept experiments in which neutral additives (e.g., Triton X-100, Tween 20, poly(ethylene glycol)) are removed from protein solutions using the MSWIFT device followed by MS analysis are described. Coupling the MSWIFT separation with ion mobility MS provides additional separation via the gas phase and assists in achieving higher quality ESI mass spectra when small amounts of additives remain in solution.
authors.affiliation: Department of Chemistry, Texas A&M University, College Station, Texas 77843, USA.
authors.forename: Pei-Jing
authors.initials: PJ
authors.lastname: Pai
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Cologna
authors.forename: William K
authors.initials: WK
authors.lastname: Russell
authors.forename: Gyula
authors.initials: G
authors.lastname: Vigh
authors.forename: David H
authors.initials: DH
authors.lastname: Russell
mesh_terms: D003902:Detergents / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D004586:Electrophoresis / Q000379:methods*; D009154:Mutation; D011506:Proteins / Q000737:chemistry*; D012996:Solutions; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics
chemical_list: D003902:Detergents; D011506:Proteins; D012996:Solutions; D013482:Superoxide Dismutase